GENE
Genetic Engineering in Education
The Genetic Engineering in Education (GENE) project enables Year 12 biology students to engage with cell biology techniques used in laboratories around the world when investigating gene function and the proteins they code for. In GENE, we work with the Green Fluorescent Protein (GFP) gene that codes for a protein that emits green light when exposed to blue or UV light. This protein, first isolated from the jellyfish Aequorea Victoria, is widely used in cell biology research as a marker for monitoring gene transfer and expression. In the GENE project, students use a range of techniques to clone the gene into a new recombinant plasmid, transform bacteria with the new plasmid and then express and recover the GFP protein from cell lysates.
In addition to our internal research sessions on Wednesday afternoons, students also attend three workshop days at the University of Kent during the year where they get hands-on experience a range of techniques involved in purifying and using proteins for diagnostics. Having recovered and purified antibody fragments expressed from a recombinant plasmid, students then chemically label these antibodies to produce a diagnostic test similar to those used in COVID-19 lateral flow test kits.
GENE Workflow
- The GFP gene is amplified by PCR from template DNA.
- Successful amplification is established by agarose gel electrophoresis.
- The PCR product, containing the GFP gene, is purified and polished ready for ligation.
- Blunt-end ligation is used to introduce the PCR product into the pJET1.2/blunt cloning vector.
- An coli strain is transformed using the new recombinant plasmid.
- Plasmids are recovered from cultured bacterial lysates using a miniprep protocol.
- Successful ligation is investigated via restriction digests, colony PCR and gene sequencing, the latter being performed using an external laboratory service.
- GFP is recovered from cell lysates and purified by Hydrophobic Interaction Chromatography (HIC).
- Correct GFP molecular weight is confirmed using SDS-PAGE electrophoresis.
- Students also get to use Benchling, a cloud-based biotech platform, which enables them to track the expected outcome from each stage of the workflow.
Bacterial colonies expressing the GFP protein, the protein fluorescing green when exposed to a UV light source.
The GENE project is generously supported by the Langton PTFA, which provides annual funding to ensure the continuation of this valuable extracurricular programme. Their support enables students to gain hands-on experience with advanced molecular biology techniques, preparing them for further studies and careers in biosciences.